@article{benedict_parsley_mochizuki_2024, title={What is your diagnosis? Blood smear from a Corolla wild horse stallion}, volume={2}, ISSN={["1939-165X"]}, url={https://doi.org/10.1111/vcp.13319}, DOI={10.1111/vcp.13319}, abstractNote={Veterinary Clinical PathologyEarly View WHAT IS YOUR DIAGNOSIS? What is your diagnosis? Blood smear from a Corolla wild horse stallion William Benedict, William Benedict orcid.org/0009-0000-3416-3201 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorAshley Parsley, Ashley Parsley orcid.org/0000-0002-0676-8833 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorHiroyuki Mochizuki, Corresponding Author Hiroyuki Mochizuki [email protected] orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA Correspondence Hiroyuki Mochizuki, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: [email protected]Search for more papers by this author William Benedict, William Benedict orcid.org/0009-0000-3416-3201 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorAshley Parsley, Ashley Parsley orcid.org/0000-0002-0676-8833 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorHiroyuki Mochizuki, Corresponding Author Hiroyuki Mochizuki [email protected] orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA Correspondence Hiroyuki Mochizuki, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: [email protected]Search for more papers by this author First published: 01 February 2024 https://doi.org/10.1111/vcp.13319Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. CONFLICT OF INTEREST STATEMENT The authors have no affiliations or financial involvement with any organization or entity with a financial interest in, or in financial compensation with, the subject matter or materials discussed in this article. REFERENCES 1Casiraghi M, Bain O, Guerrero R, et al. Mapping the presence of Wolbachia Pipientis on the phylogeny of filarial nematodes: evidence for symbiont loss during evolution. Int J Parasitol. 2004; 34: 191-203. 10.1016/j.ijpara.2003.10.004 PubMedWeb of Science®Google Scholar 2Casiraghi M, Anderson C, Bandi C, et al. A phylogenetic analysis of filarial nematodes: comparison with the phylogeny of Wolbachia endosymbionts. Parasitology. 2010; 122: 93-103. 10.1017/S0031182000007149 Google Scholar 3Coleman SU, Klei TR, French DD. Prevalence of Setaria equina (nematode: Onchocercidae) in southeastern Louisiana horses. Parasitology. 1985; 71: 512-513. 10.2307/3281548 CASWeb of Science®Google Scholar 4Yeargan MR, Lyons ET, Kania SA, et al. Incidental isolation of Setaria equina microfilariae in preparations of equine peripheral blood mononuclear cells. Vet Parasitol. 2009; 161: 142-145. 10.1016/j.vetpar.2008.12.020 PubMedWeb of Science®Google Scholar 5Sivajothi SB, Reddy S, Srinivas SS, Naveen J. Diagnosis and management of Equine Microfilariosis. Intas Polivet. 2018; 19: 226-227. Google Scholar Early ViewOnline Version of Record before inclusion in an issue ReferencesRelatedInformation}, journal={VETERINARY CLINICAL PATHOLOGY}, author={Benedict, William and Parsley, Ashley and Mochizuki, Hiroyuki}, year={2024}, month={Feb} }
 @article{wheatley_stowe_mochizuki_2023, title={Eosinophilic cavitary effusions in cats: 48 cases (2010-2020)}, volume={10}, ISSN={["1939-165X"]}, url={https://doi.org/10.1111/vcp.13295}, DOI={10.1111/vcp.13295}, abstractNote={Abstract}, journal={VETERINARY CLINICAL PATHOLOGY}, author={Wheatley, Meagan A. and Stowe, Devorah M. and Mochizuki, Hiroyuki}, year={2023}, month={Oct} }
 @article{held_mochizuki_2023, title={Hematologic Abnormalities and Diseases Associated with Moderate-to-Marked Basophilia in a Large Cohort of Dogs}, url={https://doi.org/10.3390/vetsci10120700}, DOI={10.3390/vetsci10120700}, abstractNote={Basophilia is a rare hematologic finding in dogs. This research aimed to describe the hematologic and clinical characteristics of dogs with moderate-to-marked basophilia. CBC reports with blood smear examinations from dogs presented to the North Carolina State University Veterinary Teaching Hospital were retrospectively reviewed for basophilia (>193 cells/µL). We classified basophilia as moderate when counts were ≥500 cells/µL and marked when they reached ≥1000 cells/µL. We compared the hematologic and clinical profiles of dogs with moderate-to-marked basophilia (the basophilia group) to those without basophilia, serving as our control group. In addition, we investigated differences between dogs with marked basophilia versus those with moderate basophilia, as well as between dogs in the basophilia group with and without concurrent eosinophilia. Diseases associated with moderate-to-marked basophilia included eosinophilic lung disease (p < 0.0001), leukemia/myeloproliferative neoplasms (p = 0.004), parasitic infection (p = 0.004), mast cell tumor (p = 0.005), and inflammatory bowel disease (p = 0.02). Overall, dogs with marked basophilia had a lower frequency of inflammatory diseases (51% vs. 70%, p = 0.009) and a higher frequency of neoplastic diseases (48% vs. 26%, p = 0.003) compared to those with moderate basophilia. In the basophilia group, concurrent eosinophilia was only seen in 36% of dogs. Dogs with concurrent eosinophilia were more often diagnosed with inflammatory diseases (77% vs. 58%, p = 0.006), with fewer diagnoses of neoplasia (19% vs. 40%, p = 0.001), compared to dogs without concurrent eosinophilia. The findings of this study offer veterinary clinicians valuable guidance in determining diagnostic priorities for dogs with moderate-to-marked basophilia.}, journal={Veterinary Sciences}, author={Held, Elizabeth and Mochizuki, Hiroyuki}, year={2023}, month={Dec} }
 @article{meichner_mccleary-wheeler_mochizuki_stokol_2022, title={Editorial: Comparative Oncology-Advances in Veterinary Molecular Oncology}, volume={8}, ISSN={["2297-1769"]}, DOI={10.3389/fvets.2021.812856}, abstractNote={EDITORIAL article Front. Vet. Sci., 12 January 2022Sec. Veterinary Experimental and Diagnostic Pathology Volume 8 - 2021 | https://doi.org/10.3389/fvets.2021.812856}, journal={FRONTIERS IN VETERINARY SCIENCE}, author={Meichner, Kristina and McCleary-Wheeler, Angela L. and Mochizuki, Hiroyuki and Stokol, Tracy}, year={2022}, month={Jan} }
 @article{allen_cullen_hawkey_mochizuki_nguyen_schechter_borst_yoder_freedman_patierno_et al._2021, title={A Zebrafish Model of Metastatic Colonization Pinpoints Cellular Mechanisms of Circulating Tumor Cell Extravasation}, volume={11}, ISSN={["2234-943X"]}, DOI={10.3389/fonc.2021.641187}, abstractNote={Metastasis is a multistep process in which cells must detach, migrate/invade local structures, intravasate, circulate, extravasate, and colonize. A full understanding of the complexity of this process has been limited by the lack of ability to study these steps in isolation with detailed molecular analyses. Leveraging a comparative oncology approach, we injected canine osteosarcoma cells into the circulation of transgenic zebrafish with fluorescent blood vessels in a biologically dynamic metastasis extravasation model. Circulating tumor cell clusters that successfully extravasated the vasculature as multicellular units were isolated under intravital imaging (n = 6). These extravasation-positive tumor cell clusters sublines were then molecularly profiled by RNA-Seq. Using a systems-level analysis, we pinpointed the downregulation of KRAS signaling, immune pathways, and extracellular matrix (ECM) organization as enriched in extravasated cells (p < 0.05). Within the extracellular matrix remodeling pathway, we identified versican (VCAN) as consistently upregulated and central to the ECM gene regulatory network (p < 0.05). Versican expression is prognostic for a poorer metastasis-free and overall survival in patients with osteosarcoma. Together, our results provide a novel experimental framework to study discrete steps in the metastatic process. Using this system, we identify the versican/ECM network dysregulation as a potential contributor to osteosarcoma circulating tumor cell metastasis.}, journal={FRONTIERS IN ONCOLOGY}, author={Allen, Tyler A. and Cullen, Mark M. and Hawkey, Nathan and Mochizuki, Hiroyuki and Nguyen, Lan and Schechter, Elyse and Borst, Luke and Yoder, Jeffrey A. and Freedman, Jennifer A. and Patierno, Steven R. and et al.}, year={2021}, month={Sep} }
 @article{mochizuki_stowe_2021, title={Hematologic and clinical characteristics of dogs with circulating macrophage-like cells}, volume={50}, ISSN={["1939-165X"]}, url={https://doi.org/10.1111/vcp.12962}, DOI={10.1111/vcp.12962}, abstractNote={Abstract}, number={1}, journal={VETERINARY CLINICAL PATHOLOGY}, publisher={Wiley}, author={Mochizuki, Hiroyuki and Stowe, Devorah M.}, year={2021}, month={Mar}, pages={37–44} }
 @article{miyamoto_yoshihara_shioya_tadaki_imamura_enseki_koike_furuya_mochizuki_2021, title={Lung sound analysis in infants with risk factors for asthma development}, url={https://doi.org/10.1002/hsr2.379}, DOI={10.1002/hsr2.379}, abstractNote={Abstract}, journal={Health Science Reports}, author={Miyamoto, Manabu and Yoshihara, Shigemi and Shioya, Hiromi and Tadaki, Hiromi and Imamura, Tomohiko and Enseki, Mayumi and Koike, Hideki and Furuya, Hiroyuki and Mochizuki, Hiroyuki}, year={2021}, month={Sep} }
 @article{marin_mochizuki_mastromauro_stowe_2021, title={What is your diagnosis? Dermal mass in a dog}, volume={51}, ISSN={0275-6382 1939-165X}, url={http://dx.doi.org/10.1111/vcp.13015}, DOI={10.1111/vcp.13015}, abstractNote={Veterinary Clinical PathologyEarly View WHAT IS YOUR DIAGNOSIS? What is your diagnosis? Dermal mass in a dog Jessica Marin, Jessica Marin orcid.org/0000-0002-2451-2612 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorHiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorMichael Mastromauro, Michael Mastromauro Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorDevorah M. Stowe, Corresponding Author Devorah M. Stowe damarks@ncsu.edu orcid.org/0000-0002-4058-2995 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA Correspondence Devorah M. Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: damarks@ncsu.eduSearch for more papers by this author Jessica Marin, Jessica Marin orcid.org/0000-0002-2451-2612 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorHiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorMichael Mastromauro, Michael Mastromauro Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USASearch for more papers by this authorDevorah M. Stowe, Corresponding Author Devorah M. Stowe damarks@ncsu.edu orcid.org/0000-0002-4058-2995 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA Correspondence Devorah M. Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: damarks@ncsu.eduSearch for more papers by this author First published: 04 November 2021 https://doi.org/10.1111/vcp.13015Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Early ViewOnline Version of Record before inclusion in an issue RelatedInformation}, number={1}, journal={Veterinary Clinical Pathology}, publisher={Wiley}, author={Marin, Jessica and Mochizuki, Hiroyuki and Mastromauro, Michael and Stowe, Devorah M.}, year={2021}, month={Nov}, pages={161–163} }
 @article{chandler_mochizuki_snyder_stowe_2021, title={What is your diagnosis? Mass in the abdomen of a cat}, volume={50}, ISSN={0275-6382 1939-165X}, url={http://dx.doi.org/10.1111/vcp.12929}, DOI={10.1111/vcp.12929}, abstractNote={Veterinary Clinical PathologyVolume 50, Issue 3 p. 465-467 WHAT IS YOUR DIAGNOSIS? What is your diagnosis? Mass in the abdomen of a cat Whitney Chandler, Whitney Chandler NCSU College of Veterinary Medicine, Raleigh, NC, USASearch for more papers by this authorHiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorAmy Snyder, Amy Snyder Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorDevorah M. Stowe, Corresponding Author Devorah M. Stowe damarks@ncsu.edu orcid.org/0000-0002-4058-2995 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Correspondence Devorah M. Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA Email: damarks@ncsu.eduSearch for more papers by this author Whitney Chandler, Whitney Chandler NCSU College of Veterinary Medicine, Raleigh, NC, USASearch for more papers by this authorHiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorAmy Snyder, Amy Snyder Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorDevorah M. Stowe, Corresponding Author Devorah M. Stowe damarks@ncsu.edu orcid.org/0000-0002-4058-2995 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Correspondence Devorah M. Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA Email: damarks@ncsu.eduSearch for more papers by this author First published: 26 July 2021 https://doi.org/10.1111/vcp.12929Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume50, Issue3September 2021Pages 465-467 RelatedInformation}, number={3}, journal={Veterinary Clinical Pathology}, publisher={Wiley}, author={Chandler, Whitney and Mochizuki, Hiroyuki and Snyder, Amy and Stowe, Devorah M.}, year={2021}, month={Jul}, pages={465–467} }
 @article{mochizuki_eaton_breuhaus_stowe_2020, title={What is your diagnosis? Transtracheal wash in a horse}, volume={49}, ISSN={["1939-165X"]}, url={https://doi.org/10.1111/vcp.12907}, DOI={10.1111/vcp.12907}, abstractNote={Veterinary Clinical PathologyVolume 49, Issue 4 p. 675-677 WHAT IS YOUR DIAGNOSIS? What is your diagnosis? Transtracheal wash in a horse Hiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorErin Eaton, Erin Eaton Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorBabetta Breuhaus, Babetta Breuhaus Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorDevorah M. Stowe, Corresponding Author Devorah M. Stowe damarks@ncsu.edu orcid.org/0000-0002-4058-2995 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Correspondence Devorah M. Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: damarks@ncsu.eduSearch for more papers by this author Hiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorErin Eaton, Erin Eaton Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorBabetta Breuhaus, Babetta Breuhaus Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USASearch for more papers by this authorDevorah M. Stowe, Corresponding Author Devorah M. Stowe damarks@ncsu.edu orcid.org/0000-0002-4058-2995 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Correspondence Devorah M. Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: damarks@ncsu.eduSearch for more papers by this author First published: 21 October 2020 https://doi.org/10.1111/vcp.12907Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume49, Issue4December 2020Pages 675-677 This article also appears in:What is your diagnosis? Virtual Issue RelatedInformation}, number={4}, journal={VETERINARY CLINICAL PATHOLOGY}, publisher={Wiley}, author={Mochizuki, Hiroyuki and Eaton, Erin and Breuhaus, Babetta and Stowe, Devorah M.}, year={2020}, month={Dec}, pages={675–677} }
 @article{tomita_mochizuki_tsuboi_ogura_igarashi_goto-koshino_takahashi_ohmi_tomiyasu_ohno_et al._2019, title={Development of canine X-chromosome inactivation pattern analysis for the detection of cell clonality by incorporating the examination of the SLIT and NTRK-like family member 4 (SLITRK4) gene}, volume={125}, ISSN={["1532-2661"]}, DOI={10.1016/j.rvsc.2019.06.004}, abstractNote={X-chromosome inactivation pattern (XCIP) analysis can be used to assess the clonality of cell populations of various origin by distinguishing the methylated X chromosome from the unmethylated X chromosome. In this study, the utility of XCIP analysis was improved by incorporating the examination of AC dinucleotide repeats in SLIT and NTRK-like family member 4 (SLITRK4) gene into the previously reported CAG repeat examination of androgen receptor (AR) gene in dogs. The rate of heterozygosity when both genes were analysed (125/150, 83.3%) was higher than AR gene examination alone (86/150, 57.3%). Blood samples from heterozygous dogs in either AC-1 or AC-2 of SLITRK4 gene were examined for the corrected inactivation allele ratio (CIAR), resulting in the determination of a reference range of CIAR <3.8 in non-neoplastic cell/tissue samples. Using this analytical method, 49% (21/43) of neoplastic tissue samples from dogs showed a CIAR >3.8, indicating the presence of a clonal population. Through the present study, the availability of canine XCIP analysis was improved by incorporating the examination of the SLITRK4 gene, providing a highly useful laboratory examination system for the detection of the clonality of various cell/tissue samples in dogs.}, journal={RESEARCH IN VETERINARY SCIENCE}, author={Tomita, A. and Mochizuki, H. and Tsuboi, M. and Ogura, I and Igarashi, H. and Goto-Koshino, Y. and Takahashi, M. and Ohmi, A. and Tomiyasu, H. and Ohno, K. and et al.}, year={2019}, month={Aug}, pages={170–175} }
 @article{fisher_levine_guy_mochizuki_breen_schal_watson_2019, title={Lack of influence by endosymbiont Wolbachia on virus titer in the common bed bug, Cimex lectularius}, volume={12}, ISSN={1756-3305}, url={http://dx.doi.org/10.1186/s13071-019-3694-2}, DOI={10.1186/s13071-019-3694-2}, abstractNote={Abstract}, number={1}, journal={Parasites & Vectors}, publisher={Springer Science and Business Media LLC}, author={Fisher, Michael L. and Levine, Jay F. and Guy, James S. and Mochizuki, Hiroyuki and Breen, Matthew and Schal, Coby and Watson, David W.}, year={2019}, month={Sep} }
 @article{mochizuki_sherrick_mastromauro_stowe_2019, title={What is your diagnosis? Lymphocytes engulfing erythrocytes in a cat}, volume={48}, ISSN={0275-6382 1939-165X}, url={http://dx.doi.org/10.1111/vcp.12744}, DOI={10.1111/vcp.12744}, abstractNote={Veterinary Clinical PathologyVolume 48, Issue 4 p. 768-770 WHAT IS YOUR DIAGNOSIS? What is your diagnosis? Lymphocytes engulfing erythrocytes in a cat Hiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorEllen Sherrick, Ellen Sherrick Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorMichael Mastromauro, Michael Mastromauro Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorDevorah Marks Stowe, Corresponding Author Devorah Marks Stowe damarks@ncsu.edu Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina Correspondence Devorah Marks Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: damarks@ncsu.eduSearch for more papers by this author Hiroyuki Mochizuki, Hiroyuki Mochizuki orcid.org/0000-0002-1520-0393 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorEllen Sherrick, Ellen Sherrick Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorMichael Mastromauro, Michael Mastromauro Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorDevorah Marks Stowe, Corresponding Author Devorah Marks Stowe damarks@ncsu.edu Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina Correspondence Devorah Marks Stowe, Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, USA. Email: damarks@ncsu.eduSearch for more papers by this author First published: 28 June 2019 https://doi.org/10.1111/vcp.12744Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume48, Issue4December 2019Pages 768-770 This article also appears in:What is your diagnosis? Virtual Issue RelatedInformation}, number={4}, journal={Veterinary Clinical Pathology}, publisher={Wiley}, author={Mochizuki, Hiroyuki and Sherrick, Ellen and Mastromauro, Michael and Stowe, Devorah Marks}, year={2019}, month={Jun}, pages={768–770} }
 @article{sato_mochizuki_goto-koshino_fujiwara-igarashi_takahashi_ohno_tsujimoto_2018, title={Prognostic significance of hypermethylation of death-associated protein kinase (DAPK) gene CpG island in dogs with high-grade B-cell lymphoma.}, volume={3}, url={http://europepmc.org/abstract/med/29527805}, DOI={10.1111/vco.12395}, abstractNote={Death‐associated protein kinase (DAPK) is a serine/threonine kinase and a tumour suppressor gene. Diffuse large B‐cell lymphomas with inactivated DAPK through hypermethylation of a CpG island is known to result in a biologically aggressive phenotype in humans. This retrospective study was carried out to analyse the prognostic significance of DAPK CpG island hypermethylation in canine lymphoma. We hypothesized that DAPK CpG island hypermethylation can be a negative prognostic indicator in dogs with nodal high‐grade B‐cell lymphoma. Forty‐seven dogs with high‐grade B‐cell lymphoma, according to the updated Kiel classification, were evaluated after being treated with a CHOP (vincristine, cyclophosphamide, doxorubicin and prednisolone)‐based chemotherapy protocol. The methylation status of the DAPK CpG island was examined by methylation‐specific PCR. Progression‐free survival (PFS) and overall survival (OS) were compared using the Kaplan‐Meier analysis and log‐rank test. The cox proportional hazard regression model was used to evaluate the effect of multiple variables. Hypermethylation of the DAPK CpG island was detected in 21 of the 47 dogs. The PFS and OS in dogs with the hypermethylation (median: 220 and 266 days, respectively) were significantly shorter than those of dogs without hypermethylation (median: 301 and 412 days, respectively) (PFS, P = .036; OS, P = .007). In the multivariate analysis, hypermethylation of the DAPK CpG island remained an independent prognostic factor in predicting shortened PFS (P = .047) and OS (P = .021) as well as clinical substage b. Overall, hypermethylation of the DAPK CpG island was a negative prognostic factor in canine high‐grade B‐cell lymphoma.}, journal={Veterinary and comparative oncology}, author={Sato, M and Mochizuki, H and Goto-Koshino, Y and Fujiwara-Igarashi, A and Takahashi, M and Ohno, K and Tsujimoto, H}, year={2018}, month={Mar} }
 @article{mochizuki_motsinger-reif_bettini_moroff_breen_2017, title={Association of breed and histopathological grade in canine mast cell tumours}, volume={15}, ISSN={["1476-5829"]}, url={http://europepmc.org/abstract/med/27198171}, DOI={10.1111/vco.12225}, abstractNote={Abstract}, number={3}, journal={VETERINARY AND COMPARATIVE ONCOLOGY}, author={Mochizuki, H. and Motsinger-Reif, A. and Bettini, C. and Moroff, S. and Breen, M.}, year={2017}, month={Sep}, pages={829–839} }
 @article{mochizuki_fujiwara-igarashi_sato_goto-koshino_ohno_tsujimoto_2017, title={Genetic and epigenetic aberrations of p16 in feline primary neoplastic diseases and tumor cell lines of lymphoid and non-lymphoid origins}, volume={219}, url={https://doi.org/10.1016/j.tvjl.2016.11.017}, DOI={10.1016/j.tvjl.2016.11.017}, abstractNote={The p16 gene acts as a tumor suppressor by regulating the cell cycle and is frequently inactivated in human and canine cancers. The aim of this study was to characterize genetic and epigenetic alterations of the p16 in feline lymphoid and non-lymphoid malignancies, using 74 primary tumors and 11 tumor cell lines. Cloning of feline p16 and subsequent sequence analysis revealed 11 germline sequence polymorphisms in control cats. Bisulfite sequencing analysis of the p16 promoter region in a feline lymphoma cell line revealed that promoter methylation was associated with decreased mRNA expression. Treatment with a demethylating agent restored mRNA expression of the silenced p16. PCR amplification and sequencing analysis detected homozygous loss (five tumors, 6.7%) and a missense mutation (one tumor, 1.4%) in the 74 primary tumors analyzed. Methylation-specific PCR analysis revealed promoter methylation in 10 primary tumors (14%). Promoter methylation was frequent in B cell lymphoid tumors (7/21 tumors, 33%). These genetic and epigenetic alterations were also observed in lymphoma and mammary gland carcinoma cell lines, but not detected in non-neoplastic control specimens. These data indicate that molecular alterations of the p16 locus may be involved in the development of specific types of feline cancer, and warrant further studies to evaluate the clinical value of this evolutionarily-conserved molecular alteration in feline cancers.}, journal={The Veterinary Journal}, publisher={Elsevier BV}, author={Mochizuki, H. and Fujiwara-Igarashi, A. and Sato, M. and Goto-Koshino, Y. and Ohno, K. and Tsujimoto, H.}, year={2017}, month={Jan}, pages={27–33} }
 @article{mochizuki_thomas_moroff_breen_2017, title={Genomic profiling of canine mast cell tumors identifies DNA copy number aberrations associated with KIT mutations and high histological grade}, volume={25}, ISSN={["1573-6849"]}, url={http://europepmc.org/abstract/med/28058543}, DOI={10.1007/s10577-016-9543-7}, abstractNote={Mast cell tumor (MCT) is the most common skin malignancy of domestic dogs and presents with a widely variable clinical behavior. Although activating KIT mutations are present in approximately 20% of canine MCTs, molecular etiology is largely unknown for the majority of this cancer. Characterization of genomic alterations in canine MCTs may identify genomic regions and/or genes responsible for their development and progression, facilitating the discovery of new therapeutic targets and improved clinical management of this heterogeneous cancer. We performed genome-wide DNA copy number analysis of 109 primary MCTs derived from three popular canine breeds (the Boxer, Labrador Retriever, and Pug) as well as nontarget breeds using oligonucleotide array comparative genomic hybridization (oaCGH). We demonstrated a stepwise accumulation of numerical DNA copy number aberrations (CNAs) as tumor grade increases. DNA sequencing analysis revealed that KIT mutations were found less frequently in the Pug tumors and were strongly associated with high histological grade. Tumors with KIT mutations showed genome-wide aberrant copy number profiles, with frequent CNAs involving genes in the p53 and RB pathways, whereas CNAs were very limited in tumors with wild-type KIT. We evaluated the presence of four CNAs to predict aggressive tumor phenotypes. This approach predicted aggressive tumors with a sensitivity of 78-94% and specificity of 88-93%, when using oaCGH and droplet digital PCR platforms. Further investigation of genome regions identified in this study may lead to the development of a molecular tool for classification and prognosis, as well as identification of therapeutic target molecules.}, number={2}, journal={CHROMOSOME RESEARCH}, author={Mochizuki, Hiroyuki and Thomas, Rachael and Moroff, Scott and Breen, Matthew}, year={2017}, month={Jun}, pages={129–143} }
 @article{mochizuki_breen_2017, title={Sequence analysis of RAS and RAF mutation hot spots in canine carcinoma}, volume={15}, ISSN={["1476-5829"]}, url={https://doi.org/10.1111/vco.12275}, DOI={10.1111/vco.12275}, abstractNote={Abstract}, number={4}, journal={VETERINARY AND COMPARATIVE ONCOLOGY}, publisher={Wiley-Blackwell}, author={Mochizuki, H. and Breen, M.}, year={2017}, month={Dec}, pages={1598–1605} }
 @article{mochizuki_shapiro_breen_pathology_2016, title={Detection of Copy Number Imbalance in Canine Urothelial Carcinoma With Droplet Digital Polymerase Chain Reaction}, volume={53}, ISSN={["1544-2217"]}, url={http://europepmc.org/abstract/med/26574558}, DOI={10.1177/0300985815614975}, abstractNote={ Urothelial carcinoma (UC) is the most common neoplasm of the canine urinary tract. Clinical presentation of UC is shared with several other, more common urinary tract disorders, and this often delays diagnosis of the UC. Definitive diagnosis of UC requires histopathologic examination of a biopsy specimen, but the cost and invasiveness for these diagnostic tests often result in most diagnoses being made on the basis of clinical findings, diagnostic imaging, and cytologic examination of urine sediment. Regardless of the diagnostic process used, most UCs currently are not diagnosed until they are at an advanced clinical stage and so are associated with poor prognosis. Improved methods for earlier and less invasive detection are needed. In a previous study, the authors demonstrated the presence of highly recurrent DNA copy number aberrations (CNAs) in canine UC and hypothesized that detection of these CNAs in tumor cells can be used as a molecular diagnostic for UC. In this study, a multiplexed droplet digital polymerase chain reaction (ddPCR) assay was detected to detect and quantify CNAs of specific regions of canine chromosomes 8, 13, 19, and 36. The assay was effective at differentiating 31 neoplastic and 25 nonneoplastic bladder tissues based on copy number, with 100% sensitivity and specificity in tissue samples. CNAs were also detected by ddPCR in 67% (12 of 18) of urine DNA specimens derived from UC patients. The findings show that ddPCR is a useful molecular technique to detect CNAs and may be used as a noninvasive molecular diagnostic test for canine UC. }, number={4}, journal={VETERINARY PATHOLOGY}, author={Mochizuki, Hiroyuki and Shapiro, S.G. and Breen, M. and pathology, Veterinary}, year={2016}, month={Jul}, pages={764–772} }
 @article{mochizuki_kennedy_shapiro_breen_2015, title={BRAF Mutations in Canine Cancers}, volume={10}, ISSN={["1932-6203"]}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000355955300139&KeyUID=WOS:000355955300139}, DOI={10.1371/journal.pone.0129534}, abstractNote={Activating mutations of the BRAF gene lead to constitutive activation of the MAPK pathway. Although many human cancers carry the mutated BRAF gene, this mutation has not yet been characterized in canine cancers. As human and canine cancers share molecular abnormalities, we hypothesized that BRAF gene mutations also exist in canine cancers. To test this hypothesis, we sequenced the exon 15 of BRAF, mutation hot spot of the gene, in 667 canine primary tumors and 38 control tissues. Sequencing analysis revealed that a single nucleotide T to A transversion at nucleotide 1349 occurred in 64 primary tumors (9.6%), with particularly high frequency in prostatic carcinoma (20/25, 80%) and urothelial carcinoma (30/45, 67%). This mutation results in the amino acid substitution of glutamic acid for valine at codon 450 (V450E) of canine BRAF, corresponding to the most common BRAF mutation in human cancer, V600E. The evolutional conservation of the BRAF V600E mutation highlights the importance of MAPK pathway activation in neoplasia and may offer opportunity for molecular diagnostics and targeted therapeutics for dogs bearing BRAF-mutated cancers.}, number={6}, journal={PLOS ONE}, author={Mochizuki, Hiroyuki and Kennedy, Katherine and Shapiro, Susan G. and Breen, Matthew}, year={2015}, month={Jun} }
 @article{goto-koshino_mochizuki_sato_nakashima_hiyoshi_fujiwara-igarashi_maeda_nakamura_uchida_fujino_et al._2015, title={Construction of a multicolor GeneScan analytical system to detect clonal rearrangements of immunoglobulin and T cell receptor genes in canine lymphoid tumors}, volume={165}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000355025000009&KeyUID=WOS:000355025000009}, DOI={10.1016/j.vetimm.2015.03.005}, abstractNote={Polymerase chain reaction (PCR) amplification to detect immunoglobulin heavy chain (IgH) and T cell receptor γ-chain (TCRγ) gene rearrangements has recently become widely used as part of the diagnostic strategy for lymphoid tumors in dogs. In this study, we constructed a multicolor GeneScan analytical system to improve the sensitivity and resolution of the clonality analysis of antigen receptor gene rearrangements in dogs. We used 7 reactions per sample, with 2 PCR conditions, to amplify IgH/TCRγ and control genes. By using multicolor-labeled primers, these 7 PCR products could be combined into 3 tubes before capillary electrophoresis. Clonal rearrangement of the IgH/TCRγ genes was detected in 93.3% of dogs with multicentric lymphoma and 84.6% of dogs with gastrointestinal lymphoma. Detection sensitivity of the clonally expanded cells in the background of normal peripheral blood mononuclear cells was 1–10%. The multicolor GeneScan analytical system developed here may prove to be helpful for the diagnosis of lymphoid tumors in dogs.}, number={1-2}, journal={Veterinary Immunology and Immunopathology}, author={Goto-Koshino, Yuko and Mochizuki, Hiroyuki and Sato, Masahiko and Nakashima, Ko and Hiyoshi, Saaya and Fujiwara-Igarashi, Aki and Maeda, Shingo and Nakamura, Kenji and Uchida, Kazuyuki and Fujino, Yasuhito and et al.}, year={2015}, pages={81–87} }
 @article{mochizuki_goto-koshino_takahashi_fujino_ohno_tsujimoto_2015, title={Demonstration of the Cell Clonality in Canine Hematopoietic Tumors by X-chromosome Inactivation Pattern Analysis}, volume={52}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000347300600008&KeyUID=WOS:000347300600008}, DOI={10.1177/0300985814528217}, abstractNote={ X-chromosome inactivation pattern (XCIP) analysis has been widely used to assess cell clonality in various types of human neoplasms. In this study, a polymerase chain reaction–based canine XCIP analysis of the androgen receptor ( AR) gene was applied for the assessment of cell clonality in canine hematopoietic tumors. This XCIP analysis is based on the polymorphic CAG repeats in the AR gene and the difference of methylation status between active and inactive X chromosomes. We first examined the polymorphisms of 2 CAG tandem repeats in the AR gene in 52 male and 150 female dogs of various breeds. The 2 polymorphic CAG repeats contained 9 to 12 and 10 to 14 CAGs in the first and second CAG repeats, respectively. Of the 150 female dogs, 74 (49.3%) were heterozygous for the first and/or second polymorphic CAG tandem repeats, indicating the utility of XCIP analysis in these dogs. Canine XCIP analysis was then applied to clinical samples from female dogs with canine high-grade lymphoma, chronic myelogenous leukemia, acute myelogenous leukemia, and benign lymph node hyperplasia. Of 10 lymphoma cell samples, 9 (90%) showed skewed XCIPs, indicating their clonal origins, whereas all the nonneoplastic lymph node samples showed balanced XCIPs. Moreover, bone marrow specimen from a dog with acute myelogenous leukemia and peripheral leukocyte specimens from 2 dogs with chronic myelogenous leukemia showed skewed XCIPs. XCIP analysis was successfully employed to demonstrate the cell clonality of canine hematopoietic tumors in this study and will be applicable to evaluate the clonality in various proliferative disorders in dogs. }, number={1}, journal={Veterinary Pathology}, author={Mochizuki, H. and Goto-Koshino, Y. and Takahashi, M. and Fujino, Y. and Ohno, K. and Tsujimoto, H.}, year={2015}, pages={61–69} }
 @article{mochizuki_shapiro_breen_2015, title={Detection of BRAF Mutation in Urine DNA as a Molecular Diagnostic for Canine Urothelial and Prostatic Carcinoma}, volume={10}, ISSN={["1932-6203"]}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000366903300038&KeyUID=WOS:000366903300038}, DOI={10.1371/journal.pone.0144170}, abstractNote={Urothelial carcinoma (UC) of the lower urinary tract and prostatic carcinoma (PC) are aggressive genitourinary cancers in dogs, characterized by invasion to surrounding tissues and high metastatic potential. Current diagnosis of canine UC and PC requires histopathological examination of a biopsy. Such specimens require specialized medical equipment and are invasive procedures, limiting the availability of diagnosis by histopathology for many canine patients. Access to a non-invasive means to confirm diagnosis is currently an unmet need. Recently, the canine BRAF V595E mutation was detected in ~80% of canine UCs and PCs. In this study, we developed a droplet digital PCR (ddPCR) assay for detection of the canine BRAF V595E mutation in canine urogenital tumors. The assay was evaluated in DNA samples prepared from biopsy specimens of UC (n = 48) and PC (n = 27), as well and non-neoplastic bladder epithelium (n = 38). In addition the assay was assessed for use with DNA isolated from free catch urine samples derived from canine patients with UC (n = 23), PC (n = 3), as well as from dogs with cystitis and healthy controls (n = 37). In all cases the sensitivity to detect the mutant allele was compared with conventional Sanger sequencing. ddPCR had superior sensitivity for detection of the V595E mutation: 75% of UC, 85% of PC, and 0% of control samples were mutation positive, respectively, and the V595E mutation was detected at a level as low as just 1 in 10,000 alleles (~0.01%). Furthermore, the ddPCR assay identified the mutation in free catch urine samples from 83% of canine UC and PC patients, demonstrating its utility as a non-invasive means of diagnosis. We have shown that ddPCR is a sensitive molecular technique with the potential to facilitate accurate and non-invasive means of canine UC and PC diagnosis.}, number={12}, journal={PLOS ONE}, author={Mochizuki, Hiroyuki and Shapiro, Susan G. and Breen, Matthew}, year={2015}, month={Dec} }
 @article{mochizuki_seki_nakahara_tomita_takahashi_fujino_ohno_tsujimoto_2014, title={Chronic myelogenous leukaemia with persistent neutrophilia, eosinophilia and basophilia in a cat}, volume={16}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000336791100011&KeyUID=WOS:000336791100011}, DOI={10.1177/1098612X13505576}, abstractNote={ Chronic myelogenous leukaemia was diagnosed in a 7-year-old male neutered domestic shorthair cat. Leukocytosis (74,900/µl) – mature neutrophilia, eosinophilia and basophilia – was observed. Bone marrow aspiration revealed hypercellularity with proliferation of cells of myeloid lineage. An underlying condition leading to leukocytosis was not identified. The severe leukocytosis did not respond to antibiotic therapy. Based on these findings, chronic myelogenous leukaemia was diagnosed. Because of the absence of clinical signs, the cat was monitored without treatment until 7 months after diagnosis, when it developed pruritic skin lesions. Pruritus was controlled with oral prednisolone. Forty-two months after diagnosis, the cat developed nasal lymphoma, which was treated with radiation therapy, resulting in complete remission. The cat was still in good physical condition 63 months after diagnosis, despite the persistence of marked neutrophilia, eosinophilia and basophilia. }, number={6}, journal={Journal of Feline Medicine and Surgery}, author={Mochizuki, Hiroyuki and Seki, Takahiro and Nakahara, Yoshitaka and Tomita, Akitada and Takahashi, Masashi and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2014}, pages={517–521} }
 @article{sato_mochizuki_goto-koshino_fujiwara-igarashi_takahashi_fujino_ohno_tsujimoto_2014, title={Hypermethylation of the death-associated protein kinase CpG island in canine B-cell lymphoid tumors}, volume={161}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000343360500011&KeyUID=WOS:000343360500011}, DOI={10.1016/j.vetimm.2014.08.011}, abstractNote={Death-associated protein kinase (DAPK) is a 160-kD serine/threonine kinase known as a key molecule in interferon-γ (IFN-γ)-induced apoptosis and tumor suppression. Hypermethylation of the CpG island in DAPK inactivates the gene in a variety of human malignancies. This study aimed to detect the inactivation of DAPK in canine lymphoid tumor cells. The sequence of canine DAPK cDNA was obtained from normal dog peripheral blood mononuclear cells after reverse transcription polymerase chain reaction (RT-PCR). By rapid amplification of 5′-cDNA ends, the transcription initiation site of the DAPK gene was identified. The CpG island located upstream of the translation initiation site was identified by using a search algorithm. The methylation status of the CpG island was examined using bisulfite sequence analysis and methylation-specific PCR (MSP). The inactivation of DAPK gene was examined in 3 canine lymphoid tumor cell lines, GL-1 (B-cell leukemia), CLBL-1 (B-cell lymphoma), and CL-1 (T-cell lymphoma). DAPK mRNA expression was measured by real-time RT-PCR. IFN-γ-induced apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. The influence of demethylation was examined with 5-aza-2′-deoxycytidine (5-aza-dC). The methylation status in 14 dogs with various lymphoid tumors was screened by MSP. A 1926-bp CpG island containing 280 CpG repeats was identified upstream of the translation start site of canine DAPK. Bisulfate sequence analysis and MSP revealed hypermethylation of the CpG island in GL-1 cells, but not in CLBL-1 or CL-1 cells. The amount of DAPK mRNA was significantly smaller in GL-1 cells than CLBL-1 and CL-1 cells. IFN-γ-induced apoptosis was detected in CLBL-1 and CL-1 cells but not in GL-1 cells. Treatment with 5-aza-dC significantly increased the amount of DAPK mRNA and IFN-γ-induced apoptosis in GL-1 cells. These results revealed the inactivation of DAPK through methylation of its CpG island in GL-1 cells. MSP showed hypermethylation of the DAPK CpG island in 5 of 8 primary B-cell lymphoma samples, but not in any of the 6 primary T-cell lymphoid tumor samples obtained from canine patients. DAPK was inactivated through hypermethylation of its CpG island in canine B-cell lymphoid tumor cells. This study will lead to the use of canine B-cell lymphoid tumors as an animal model to evaluate the efficacy of demethylating agents.}, number={3-4}, journal={Veterinary Immunology and Immunopathology}, author={Sato, Masahiko and Mochizuki, Hiroyuki and Goto-Koshino, Yuko and Fujiwara-Igarashi, Aki and Takahashi, Masashi and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2014}, pages={222–231} }
 @article{fujiwara-igarashi_goto-koshino_mochizuki_sato_fujino_ohno_tsujimoto_2014, title={Inhibition of p16 tumor suppressor gene expression via promoter hypermethylation in canine lymphoid tumor cells}, volume={97}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000340852600011&KeyUID=WOS:000340852600011}, DOI={10.1016/j.rvsc.2014.04.008}, abstractNote={To investigate the epigenetic regulation of the p16 gene in canine lymphoid tumor cells, its methylation status was examined in four canine lymphoid tumor cell lines. In three canine lymphoid tumor cell lines (CLBL-1, GL-1, and UL-1) with low-level p16 mRNA expression, 20 CpG sites in the promoter region of p16 gene were consistently methylated although all of the CpG sites were not methylated in another cell line (CL-1) and normal lymph node cells. The expression level of p16 mRNA in these three cell lines was restored after cultivation in the presence of a methylation inhibitor, 5-Aza-2′-deoxycitidine, indicating inactivation of p16 gene via hypermethylation. This study revealed the inactivation of p16 gene through hypermethylation of its CpG island in a fraction of canine lymphoid tumor cells.}, number={1}, journal={Research in Veterinary Science}, author={Fujiwara-Igarashi, Aki and Goto-Koshino, Yuko and Mochizuki, Hiroyuki and Sato, Masahiko and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2014}, pages={60–63} }
 @article{fujiwara-igarashi_goto-koshino_mochizuki_maeda_fujino_ohno_tsujimoto_2013, title={Simultaneous Inactivation of the p16, p15 and p14 Genes Encoding Cyclin-Dependent Kinase Inhibitors in Canine T-Lymphoid Tumor Cells}, volume={75}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000330268300008&KeyUID=WOS:000330268300008}, DOI={10.1292/jvms.12-0351}, abstractNote={The p16, p15 and p14 genes are widely known as tumor suppressor genes in human medicine. Although a large number of genetic and epigenetic aberrations in these genes have been reported in human malignancies, canine malignancies have not been well analyzed on the aberrations of these genes. In this study, the full-length complementary DNA (cDNA) of the canine p16 gene was cloned using the 5' and 3' rapid amplification of cDNA ends methods. Based on the sequence data, primers specific for p16, p15 and p14 were designed. Using these primers, the expression of p16, p15 and p14 mRNAs could be individually evaluated by reverse transcriptase polymerase chain reaction. Genomic aberrations were also examined using genomic polymerase chain reaction. Two of the 6 canine lymphoid tumor cell lines did not express detectable levels of p16, p15 and p14 mRNAs, and wide-ranging deletions in the p15-p14-p16 genomic locus were suspected. Wide-ranging deletions were also speculated in 2 of 14 dogs with T-cell lymphoid tumors. On the other hand, similar failure of amplification suggesting wide-ranging deletions were not observed in any of the 14 dogs with B-cell lymphoma. Deletion of the p15-p14-p16 genomic locus could be one of the molecular aberrations in canine lymphoid tumor cells.}, number={6}, journal={Journal of Veterinary Medical Science}, author={Fujiwara-Igarashi, Aki and Goto-Koshino, Yuko and Mochizuki, Hiroyuki and Maeda, Shingo and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2013}, pages={733–742} }
 @article{mochizuki_goto-koshino_sato_fujino_ohno_tsujimoto_2012, title={Comparison of the antitumor effects of an MDM2 inhibitor, nutlin-3, in feline lymphoma cell lines with or without p53 mutation}, volume={147}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000305435100009&KeyUID=WOS:000305435100009}, DOI={10.1016/j.vetimm.2012.04.017}, abstractNote={The P53 tumor suppressor protein is a multifunctional transcription factor that prevents the malignant transformation of normal cells. In human malignancies, p53 is the most frequently altered gene and is mutated in approximately 50% of all malignancies. In contrast, p53 gene mutation has been rarely detected in feline malignancies, and most feline malignancies conceivably retain the wild-type p53 (wt-p53) gene. MDM2 negatively regulates the P53 protein by inhibiting its transcriptional activity and nuclear transport and by inducing its degradation. Inhibition of P53–MDM2 interaction stabilizes P53 protein and activates P53 pathway. Nutlin-3, a small molecule that inhibits P53–MDM2 interaction, was shown to have an antitumor effect in several human cancer cells retaining the wt-p53 gene. In the present study, we evaluated and compared the antitumor effect of nutlin-3 in 5 different feline lymphoma cell lines, of which 3 harbored wt-p53, and 2, mutated p53 (mt-p53). Treatment with nutlin-3 resulted in increased amounts of P53 protein in conjunction with augmented expression of P53-target genes in 3 feline lymphoma cell lines with the wt-p53 gene, but not in 2 feline lymphoma cell lines with the mt-p53 gene. Nutlin-3 treatment also induced G1-S and/or G2-M cell cycle arrest and apoptosis in lymphoma cell lines with wt-p53. Nutlin-3 treatment induced cell cycle arrest but not apoptosis in the cell lines with mt-p53. From these results, we concluded that nutlin-3 has an antitumor effect on feline lymphoma cell lines harboring the wt-p53 gene through accumulation and activation of P53 leading to cell cycle arrest and apoptosis. The present study suggests that inhibition of P53–MDM2 interaction using nutlin-3 may be a new therapeutic strategy for treating feline lymphoma retaining the wt-p53 gene.}, number={3-4}, journal={Veterinary Immunology and Immunopathology}, author={Mochizuki, Hiroyuki and Goto-Koshino, Yuko and Sato, Masahiko and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2012}, pages={187–194} }
 @article{sato_kanemoto_kagawa_kobayashi_goto-koshino_mochizuki_takahashi_fujino_ohno_tsujimoto_2012, title={Evaluation of the prognostic significance of BCL6 gene expression in canine high-grade B-cell lymphoma}, volume={191}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000300340700019&KeyUID=WOS:000300340700019}, DOI={10.1016/j.tvjl.2010.12.006}, abstractNote={The clinical usefulness of BCL6 gene expression was evaluated as a prognostic indicator in dogs with high-grade B-cell lymphoma. Forty-four dogs were diagnosed with centroblastic or B-cell immunoblastic type lymphoma according to the updated Kiel classification. BCL6 mRNA expression was measured by real-time PCR and its relationship with prognosis was analyzed. Progression-free and overall survival was not significantly different between the high BCL6 expression group (higher than the median) and the low BCL6 expression group (lower than the median) (P=0.99 and P=0.61, respectively). No correlation between BCL6 and prognosis was observed in this study, which is inconsistent with findings reported for human diffuse large B-cell lymphoma. BCL6 protein expression was not detected in the 11 dogs evaluated by immunohistochemistry. Furthermore, BCL6 protein expression was assessed in 13 archived paraffin-embedded high-grade canine lymphoma tissues and all were also negative. The results suggest that most canine high-grade B-cell lymphomas correspond to human diffuse large B-cell lymphoma with no immunohistochemical expression of BCL6.}, number={1}, journal={Veterinary Journal}, author={Sato, Masahiko and Kanemoto, Hideyuki and Kagawa, Yumiko and Kobayashi, Tetsuya and Goto-Koshino, Yuko and Mochizuki, Hiroyuki and Takahashi, Masashi and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2012}, pages={108–114} }
 @article{mochizuki_nakamura_sato_goto-koshino_sato_takahashi_fukushima_nakashima_fujino_ohno_et al._2012, title={GeneScan analysis to detect clonality of T-cell receptor gamma gene rearrangement in feline lymphoid neoplasms}, volume={145}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000301159200046&KeyUID=WOS:000301159200046}, DOI={10.1016/j.vetimm.2011.12.015}, abstractNote={Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. Polymerase chain reaction (PCR) amplification of the complementarity-determining region (CDR) 3 of the T-cell receptor (TCR) γ gene can be used to assess clonality of T-cell populations as a supportive diagnostic tool for T-cell neoplasms. Because the length variation in the TCRγ CDR3 is relatively small, false positive results may occur in non-neoplastic T-cell populations in the absence of high-resolution analytical methods for PCR products. In the present study, a PCR assay system was developed to detect clonal TCRγ gene rearrangement in feline lymphoid cells using GeneScan analysis. Thirty T-cell neoplasms, 27 B-cell neoplasms, and 34 non-neoplastic tissues were subjected to the newly developed TCRγ gene rearrangement analysis. Clonal TCRγ gene rearrangement was detected in 26 of 30 (87%) T-cell neoplasms, 2 of 27 (7%) B-cell neoplasms, and 1 of 34 (3%) non-neoplastic tissues. To compare GeneScan analysis with conventional PAGE and heteroduplex analysis, 20 clonal and 20 polyclonal samples were subjected to both analyses. Most of the results were concordant between the 2 analyses; however, several clonal peaks (bands) appeared as a single band when analyzed via conventional PAGE with heteroduplex analysis in 4 of the 20 (20%) clonal samples as a result of the difference in resolution. The PCR assay system to detect clonal TCRγ gene rearrangement in feline lymphoid cells, using GeneScan analysis, would be a useful molecular diagnostic tool for feline T-cell neoplasms, with high fidelity.}, number={1-2}, journal={Veterinary Immunology and Immunopathology}, author={Mochizuki, Hiroyuki and Nakamura, Kenji and Sato, Hirofumi and Goto-Koshino, Yuko and Sato, Masahiko and Takahashi, Masashi and Fukushima, Kenjiro and Nakashima, Ko and Fujino, Yasuhito and Ohno, Koichi and et al.}, year={2012}, pages={402–409} }
 @article{mochizuki_goto-koshino_takahashi_fujino_ohno_tsujimoto_2012, title={X-Chromosome Inactivation Pattern Analysis for the Assessment of Cell Clonality in Cats}, volume={49}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000310853200010&KeyUID=WOS:000310853200010}, DOI={10.1177/0300985812439216}, abstractNote={ X-chromosome inactivation pattern (XCIP) analysis has been widely used to assess cell clonality in various types of neoplasms in humans. In the present study, a polymerase chain reaction–based feline XCIP analysis using the feline androgen receptor gene was developed. To construct the system of the analysis, polymorphism in CAG tandem repeats within the feline androgen receptor gene was explored using somatic DNAs from 50 male and 103 female cats. CAG tandem repeats in exon 1 of the feline androgen receptor gene were found to be polymorphic, containing 15 to 22 CAG repeats. Of the 103 female cats, 70 (68%) were heterozygous for the number of CAG repeats, indicating the possible usefulness of XCIP analysis in cats. Application of the feline XCIP analysis to 3 feline mammary gland adenocarcinoma cell lines revealed distinctly skewed XCIPs in these cell lines, indicating their clonal origins. Twelve (80%) of the 15 primary tissue/cell samples obtained from cats with various neoplastic diseases showed skewed XCIPs. Moreover, bone marrow samples from 3 cats with myelodysplastic syndrome were also found to have skewed XCIPs. The polymerase chain reaction–based XCIP analysis developed in this study can provide information on cell clonality in female cats, potentially facilitating the differential diagnosis of various disorders in cats. }, number={6}, journal={Veterinary Pathology}, author={Mochizuki, H. and Goto-Koshino, Y. and Takahashi, M. and Fujino, Y. and Ohno, K. and Tsujimoto, H.}, year={2012}, pages={963–970} }
 @article{goto-koshino_ohno_nakajima_mochizuki_kanemoto_tsujimoto_2011, title={A Rapid and Simple Method to Obtain Canine Peripheral Blood-Derived Macrophages}, volume={73}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000293043900010&KeyUID=WOS:000293043900010}, DOI={10.1292/jvms.10-0470}, abstractNote={Macrophages play an important role in a variety of situations, including pathogen elimination, inflammation, and tissue repair. However, these cells are not fully studied in dogs, in part, due to the difficulty of efficiently isolating and culturing them in vitro. In this study, we cultured canine peripheral blood mononuclear cells (PBMCs) with 10 ng/ml of phorbol 12-myristate-13-acetate (PMA) for 5 days to obtain macrophages. A high number of round-adherent cells were obtained without the addition of any cytokine. These cells showed active phagocytic activity and a cell surface antigen profile different from dendritic cells. Our method facilitates a high yield of macrophages in a short cultivation period compared with previous studies. This method might be a powerful tool to study macrophage functions in dogs.}, number={6}, journal={Journal of Veterinary Medical Science}, author={Goto-Koshino, Yuko and Ohno, Koichi and Nakajima, Mayumi and Mochizuki, Hiroyuki and Kanemoto, Hideyuki and Tsujimoto, Hajime}, year={2011}, pages={773–778} }
 @article{sato_goto-koshino_kanemoto_mochizuki_fujino_ohno_tsujimoto_2011, title={Characterization of canine BCL6 cDNA and detection systems for its protein expression}, volume={140}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000288342800021&KeyUID=WOS:000288342800021}, DOI={10.1016/j.vetimm.2010.08.009}, abstractNote={BCL6 is known to be a key molecule in germinal center (GC) formation of lymph nodes, and its expression profiles have been implicated in the prognosis of diffuse large B-cell lymphoma in humans. The present study was carried out to characterize canine BCL6 cDNA and to indicate the technical methods for detection of the BCL6 protein in dog tissues. The deduced amino acid sequence of canine BCL6 showed close homology to that of human BCL6 (96.3%), especially in the zinc-finger motifs and POZ (poxvirus and zinc finger) domain with complete identity. Immunoblot analysis of a canine lymph node with an anti-human BCL6 monoclonal antibody revealed a band of 80 kDa. Immunohistochemical staining using the same antibody produced positive reactions in the cells exclusively localized in the GC of a canine lymph node. This study will be useful for the molecular classification of canine B-cell lymphomas with different prognoses.}, number={1-2}, journal={Veterinary Immunology and Immunopathology}, author={Sato, Masahiko and Goto-Koshino, Yuko and Kanemoto, Hideyuki and Mochizuki, Hiroyuki and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2011}, pages={166–169} }
 @article{mochizuki_takahashi_nishigaki_ide_goto-koshino_watanabe_sato_sato_kotera_fujino_et al._2011, title={Establishment of a novel feline leukemia virus (FeLV)-negative B-cell cell line from a cat with B-cell lymphoma}, volume={140}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000289139600018&KeyUID=WOS:000289139600018}, DOI={10.1016/j.vetimm.2010.12.010}, abstractNote={We established a novel feline B-cell line, MS4, from the neoplastic pleural effusion of a cat with cutaneous B-cell lymphoma. Immunophenotype staining of the MS4 cells was positive for CD20, CD79α, and IgA and negative for CD3, CD4, CD5, CD8α, CD18, CD21, CD22, IgM, IgG, Ig light chain, and MHC class II. PCR analysis for immunoglobulin heavy chain gene rearrangements revealed a monoclonal rearrangement, whereas no clonal rearrangement of the T-cell receptor γ gene was detected. Southern blotting with an exogenous feline leukemia virus (FeLV) U3 probe revealed no integration of exogenous FeLV provirus. The MS4 cell line is the first FeLV-negative feline B-cell lymphoma cell line, and may be used to investigate the pathogenesis of spontaneously occurring feline lymphoma and the development of new therapies.}, number={3-4}, journal={Veterinary Immunology and Immunopathology}, author={Mochizuki, Hiroyuki and Takahashi, Masashi and Nishigaki, Kazuo and Ide, Tetsuya and Goto-Koshino, Yuko and Watanabe, Shinya and Sato, Hirofumi and Sato, Masahiko and Kotera, Yukiko and Fujino, Yasuhito and et al.}, year={2011}, pages={307–311} }
 @article{mochizuki_nakamura_sato_goto-koshino_sato_takahashi_fujino_ohno_uchida_nakayama_et al._2011, title={Multiplex PCR and Genescan analysis to detect immunoglobulin heavy chain gene rearrangement in feline B-cell neoplasms}, volume={143}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000294574200005&KeyUID=WOS:000294574200005}, DOI={10.1016/j.vetimm.2011.05.030}, abstractNote={Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.}, number={1-2}, journal={Veterinary Immunology and Immunopathology}, author={Mochizuki, Hiroyuki and Nakamura, Kenji and Sato, Hirofumi and Goto-Koshino, Yuko and Sato, Masahiko and Takahashi, Masashi and Fujino, Yasuhito and Ohno, Koichi and Uchida, Kazuyuki and Nakayama, Hiroyuki and et al.}, year={2011}, pages={38–45} }
 @article{sato_goto-koshino_mochizuki_fujino_ohno_tsujimoto_2011, title={Perfusion method for harvesting bone marrow cells from dogs}, volume={72}, url={http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=ORCID&SrcApp=OrcidOrg&DestLinkType=FullRecord&DestApp=WOS_CPL&KeyUT=WOS:000295390000007&KeyUID=WOS:000295390000007}, number={10}, journal={American Journal of Veterinary Research}, author={Sato, Masahiko and Goto-Koshino, Yuko and Mochizuki, Hiroyuki and Fujino, Yasuhito and Ohno, Koichi and Tsujimoto, Hajime}, year={2011}, pages={1344–1348} }